Note: Descriptions are shown in the official language in which they were submitted.
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
METHODS OF SYNTHESIZING SUBSTANTIALLY
MONODISPERSED MIXTURES OF POLYMERS HAVING
POLYETHYLENE GLYCOL MOIETIES
Field Of The Invention
The present invention relates to methods of synthesizing polymeric compounds,
and
more particularly, to methods of synthesizing polymeric compounds comprising
polyethylene
glycol moieties.
Background Of The Invention
Polyethylene glycol (PEG) is used in a wide variety of applications including,
but not
limited to, plasticizers, softeners, humectants, ointments, polishes, paper
coating, mold
lubricants, bases for cosmetics and pharmaceuticals, solvents, binders, metal
and rubber
processing, and additives to foods and animal feed. Some particular uses of
PEG in
pharmaceutical applications include, for example, formation of PEG-drug
conjugates,
treatment of neonatal respiratory distress syndrome, treatment of functional
and/or chronic
constipation, treatment of encopresis in children, and diagnosis and therapy
of
gastrointestinal diseases.
PEG is typically produced by base-catalyzed ring-opening polymerization of
ethylene
oxide. The reaction is initiated by adding ethylene oxide to ethylene glycol,
with potassium
hydroxide as catalyst. This process results in a polydispersed mixture of
polyethylene glycol
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
polymers having a molecular weight within a given range of molecular weights.
For
example, PEG products offered by Sigma-Aldrich of Milwaukee, Wisconsin are
provided in
polydispersed mixtures such as PEG 400 (Mn 380-420); PEG 1,000 (Mõ 950-1,050);
PEG
1,500 (Mn 1,400-1,600); and PEG 2,000 (Mõ 1,900-2,200).
In J. Milton Harris, Laboratory Synthesis of Polyethylene Glycol Derivatives,
25(3)
Rev. Macromol. Chem. Phys. 325-373 (1985), the author discusses synthesis of
monomethyl
ethers of PEG (also known as methyl-terminated PEG or mPEG). The reference
states that
mPEG contains a significant amount (as much as 25%; from size exclusion
chromatography)
of PEG without the methoxy end group. This PEG "impurity" may result from
water present
in the polymerization process. Under basic conditions, hydroxide is produced,
which yields
PEG upon reaction with the ethylene oxide monomer. Since the hydroxide-
initiated PEG
chain can grow at both ends, while the methoxide-initiated chain can grow from
only one
end, the resulting mixture has a broader molecular weight distribution than
that for the PEG's.
While these polydispersed mixtures of PEGs and/or mPEGs may be useful for some
applications, physical properties of polymers may vary with the length of the
polymer. Thus,
polydispersed mixtures may not be suitable for certain applications that
require specific
physical properties. Additionally, the heterogeneity of commercially available
PEGs and
mPEGs may complicate spectroscopic analysis, physico-chemical characterization
and
pharmacokinetics analysis. As a result, it is desirable to provide
monodispersed mixtures of
PEGs and/or mPEGs.
Monodispersed mixtures of PEG and/or mPEG polymers may be provided by various
organic synthesis routes. For example, in Yiyan Chen & Gregory L. Baker,
Synthesis and
Properties of ABA Amphiphiles, 64 J. Org. Chem. 6870-6873 (1999), the authors
propose the
following scheme:
2
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
H(OCH2CH2)aOH Trityl Chloride H(OCH2CH2)aOTr
Pyridine
NaH
Na(OCH2CH2)aOTr
Ts(OCH2CH2)bOTs
p-toluene sulfonyl chloride
KOH
H(OCH2CH2)bOH Tr(OCH2CH2)2a+bOTr
H2 (50 atm)
Pd/C
NaH
R(OCH2CH2)yOR H(OCH2CH2)2a+bOH
CH3(CH2)1,-,Br
This synthesis route may be inconvenient due to the number of steps required
as well as the
use of undesirable reaction conditions such as high temperatures that may
actually break
down the PEG polymer. Moreover, it may be difficult to purify the product as
the starting
material may always be present in the reaction mixture.
In Gerard Coudert et al., A Novel, Unequivocal Synthesis of Polyethylene
Glycols,
Synthetic Communications, 16(1): 19-26 (1986), the authors proposed the
following
synthesis route:
H(OC2H4)30H Benzyl chloride BZl(OC2H4)30H
50% NaOH
1) Benzyl chloride / 50% NaOH
H(OC2H4)20H B4OC2H4)2C1
2) Thionyl Chloride,0
Bzl(OC2H4)5OBzI H2 / Pd-c H(OC2H4)50H
3
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
This synthesis route may be inconvenient due to the undesirable reaction
conditions, which
do not lead to mPEG.
As a result, it is desirable to provide a new route for synthesizing PEG,
mPEG, and/or
polymers comprising a PEG moiety that are more efficient and do not require
such
undesirable reaction conditions.
Summary Of The Invention
Embodiments of the present invention provide improved methods for synthesizing
substantially monodispersed mixtures of polymers comprising polyethylene
glycol moieties.
Methods according to embodiments of the present invention may utilize reaction
conditions
that are milder than those required by the conventional methods described
above. For
example, many, if not all, of the steps of methods according to embodiments of
the present
invention may be carried out at atmospheric pressure and/or at room
temperature. The ability
to perform these steps at atmospheric pressure and/or temperature may reduce
or prevent the
formation of undesirable side products. Additionally, methods according to
embodiments of
the present invention may be more efficient than the conventional methods
described above.
For example, methods according to embodiments of the present invention may
require fewer
steps and/or less time than the conventional methods described above. Methods
according to
embodiments of the present invention may provide the ability to remove PEG
starting
materials from the products comprising polyethylene glycol moieties to provide
substantially
monodispersed mixtures of polymers comprising polyethylene glycol moieties.
According to embodiments of the present invention, a method of synthesizing a
substantially monodispersed mixture of polymers comprising polyethylene glycol
moieties
includes:
reacting a substantially monodispersed mixture of compounds having the
structure of
Formula I:
R' (OC2H4)õ-O-X+ (I)
wherein R' is H or a lipophilic moiety; n is from 1 to 25; and X+ is a
positive
ion,
with a substantially monodispersed mixture of compounds having the structure
of Formula II:
R2(OC2H4)m OMs (II)
wherein R2 is H or a lipophilic moiety; and in is from I to 25,
4
CA 02449698 2010-01-18
under conditions sufficient to provide a substantially monodispersed mixture
of polymers
comprising polyethylene glycol moieties and having the structure of Formula
III:
R2(OC2114)m+n OR' (III).
Methods according to embodiments of the present invention may provide more
efficient synthesis routes for substantially monodispersed mixtures of PEGs,
substantially
monodispersed mixtures of mPEGs and/or substantially monodispersed mixtures of
polymers comprising PEG moieties. Methods of the present invention may reduce
the
number of steps and/or reduce the overall synthesis time compared to
conventional methods
of synthesizing PEG polymers. Methods of the present invention may also
utilize milder
reaction conditions than those used in conventional methods.
In accordance with an aspect of the present invention, there is provided a
method of
synthesizing a substantially monodispersed mixture of polymers comprising
polyethylene
glycol moieties, said method comprising:
reacting a substantially monodispersed mixture of compounds having the
structure
of Formula V:
R2(OC2H4)m -OH (V)
with a methanesulfonyl halide under conditions sufficient to provide a
substantially
monodispersed mixture of compounds having the structure of Formula II:
R2(OC2H4)m-OMs (II)
wherein R2 is H or a lipophilic moiety; and in is from 1 to 25, and
reacting the compound of Formula II with a substantially monodispersed mixture
of
compounds having the structure of Formula I:
R'(OC2H )n-O-X+ (I)
wherein R1 is H or a lipophilic moiety; n is from 1 to 25; and X+ is a
positive
ion,
under conditions sufficient to provide a substantially monodispersed mixture
of polymers
comprising polyethylene glycol moieties and having the structure of Formula
III:
R2(OC2H4)m+n-ORS (III).
Brief Description of the Drawings
Figure 1 illustrates a generic scheme for synthesizing a mixture of activated
polymers comprising a polyethylene glycol moiety and a fatty acid moiety
according to
embodiments of the present invention;
5
CA 02449698 2010-01-18
Figure 2 illustrates a scheme for synthesizing a mixture of activated mPEG7-
hexyl
oligomers according to embodiments of the present invention; and
Figure 3 illustrates a scheme for synthesizing a mixture of mPEG according to
embodiments of the present invention.
Detailed Description Of Preferred Embodiments
The invention will now be described with respect to preferred embodiments
described herein. It should be appreciated however that these embodiments are
for the
purpose of illustrating the invention, and are not to be construed as limiting
the scope of the
invention as defined by the claims.
As used herein, the term "non-polydispersed" is used to describe a mixture of
compounds having a dispersity that is in contrast to the polydispersed
mixtures of PEG
products offered by Sigma-Aldrich of Milwaukee, Wisconsin such as PEG 400 (Mn
380-
420); PEG 1,000 (Mn 950-1,050); PEG 1,500 (Mn 1, 400-1,600); and PEG 2,000 (Mn
1,
900- 2,200).
As used herein, the term "substantially monodispersed" is used to describe a
mixture
of compounds wherein at least about 95 percent of the compounds in the mixture
have the
5a
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
same molecular weight.
As used herein, the term "monodispersed" is used to describe a mixture of
compounds
wherein about 100 percent of the compounds in the mixture have the same
molecular weight.
As used herein, the term "weight average molecular weight" is defined as the
sum of
the products of the weight fraction for a given molecule in the mixture times
the mass of the
molecule for each molecule in the mixture. The "weight average molecular
weight" is
represented by the symbol M.
As used herein, the term "number average molecular weight" is defined as the
total
weight of a mixture divided by the number of molecules in the mixture and is
represented by
the symbol M,,.
As used herein, the term "PEG" refers to straight or branched polyethylene
glycol
polymers, and includes the monomethylether of polyethylene glycol (mPEG). The
terms
"PEG subunit" and polyethylene glycol subunit refer to a single polyethylene
glycol unit, i.e.,
-(CH2CHZO)-.
As used herein, the term "lipophilic" means the ability to dissolve in lipids
and/or the
ability to penetrate, interact with and/or traverse biological membranes, and
the term,
"lipophilic moiety" or "lipophile" means a moiety which is lipophilic and/or
which, when
attached to another chemical entity, increases the lipophilicity of such
chemical entity.
Examples of lipophilic moieties include, but are not limited to, alkyls, fatty
acids, esters of
fatty acids, cholesteryl, adamantyl and the like.
As used herein, the term "lower alkyl" refers to substituted or unsubstituted
alkyl
moieties having from I to 5 carbon atoms.
As used herein, the term "higher alkyl" refers to substituted or unsubstituted
alkyl
moieties having 6 or more carbon atoms.
According to aspects of the present invention, a substantially monodispersed
mixture
of polymers comprising polyethylene glycol moieties is provided as illustrated
in reaction 1:
RI(OC2H4),O X+ + R2(OC2H4)mOMs R2(OC2H4)m+nOR' 1
(I) (II) (III)
R' is H or a lipophilic moiety. R' is preferably H, alkyl, aryl alkyl, an
aromatic
moiety, a fatty acid moiety, an ester of a fatty acid moiety, cholesteryl, or
adamantyl. R' is
more preferably H, lower alkyl, or an aromatic moiety. R' is most preferably
H, methyl, or
benzyl.
6
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
The value of n is from I to 25. Preferably n is from I to 6.
X+ is a positive ion. Preferably X+ is any positive ion in a compound, such as
a strong
base, that is capable of ionizing a hydroxyl moiety on PEG. Examples of
positive ions
include, but are not limited to, sodium ions, potassium ions, lithium ions,
cesium ions, and
thallium ions.
R2 is H or a lipophilic moiety. R2 is preferably branched or linear alkyl,
aryl alkyl, an
aromatic moiety, a fatty acid moiety, or an ester of a fatty acid moiety. R2
is more preferably
lower alkyl, benzyl, a fatty acid moiety having 1 to 24 carbon atoms, or an
ester of a fatty
acid moiety having 1 to 24 carbon atoms. R2 is most preferably methyl, a fatty
acid moiety
having I to 18 carbon atoms or an ethyl ester of a fatty acid moiety having 1
to 18 carbon
atoms.
The value of m is from 1 to 25. Preferably m is from 1 to 6.
Ms is a mesylate moiety (i.e., CH3S(O2)-).
As illustrated in reaction 1, a mixture of compounds having the structure of
Formula I
is reacted with a mixture of compounds having the structure of Formula II to
provide a
mixture of polymers comprising polyethylene glycol moieties and having the
structure of
Formula III. The mixture of compounds having the structure of Formula I is a
substantially
monodispersed mixture. Preferably, at least 96, 97, 98 or 99 percent of the
compounds in the
mixture of compounds of Formula I have the same molecular weight, and, more
preferably,
the mixture of compounds of Formula I is a monodispersed mixture. The mixture
of
compounds of Formula II is a substantially monodispersed mixture. Preferably,
at least 96,
97, 98 or 99 percent of the compounds in the mixture of compounds of Formula
II have the
same molecular weight, and, more preferably, the mixture of compounds of
Formula II is a
monodispersed mixture. The mixture of compounds of Formula III is a
substantially
monodispersed mixture. Preferably, at least 96, 97, 98 or 99 percent of the
compounds in the
mixture of compound of Formula III have the same molecular weight. More
preferably, the
mixture of compounds of Formula III is a monodispersed mixture.
Reaction 1 is preferably performed between about 0 C and about 40 C, is more
preferably performed between about 15 C and about 35 C, and is most preferably
performed
at room temperature (approximately 25 C).
7
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
Reaction 1 may be performed for various periods of time as will be understood
by
those skilled in the art. Reaction 1 is preferably performed for a period of
time between
about 0.25, 0.5 or 0.75 hours and about 2, 4 or 8 hours.
Reaction 1 is preferably carried out in an aprotic solvent such as, but not
limited to,
N,N-dimethylacetamide (DMA), N,N-dimethylformamide (DMF), dimethyl sulfoxide,
hexamethylphosphoric triamide, tetrahydrofuran (THF), dioxane, diethyl ether,
methyl t-butyl
ether (MTBE), toluene, benzene, hexane, pentane, N-methylpyrollidinone,
tetrahydronaphthalene, decahydronaphthalene, 1,2-dichlorobenzene, 1,3-dimethyl-
2-
imidazolidinone, or a mixture thereof. More preferably, the solvent is DMF,
DMA or
toluene.
The molar ratio of the compound of Formula Ito the compound of Formula II is
preferably greater than about 1:1. More preferably, the molar ratio is at
least about 2:1. By
providing an excess of the compounds of Formula I, one can ensure that
substantially all of
the compounds of Formula II are reacted, which may aid in the recovery of the
compounds of
Formula III as discussed below.
Compounds of Formula I are preferably prepared as illustrated in reaction 2:
compound capable of l + 30 R~(OC2Hq)nOH + ionizing a hydroxyl moiety R
(OC2H4)nO X 2
on the PEG moiety of
(IV) Formula IV (1)
R' and X+ are as described above and the mixture of compounds of Formula IV is
substantially monodispersed; preferably, at least 96, 97, 98 or 99 percent of
the compounds in
the mixture of compounds of Formula IV have the same molecular weight; and,
more
preferably, the mixture of compounds of Formula IV is a monodispersed mixture.
Various compounds capable of ionizing a hydroxyl moiety on the PEG moiety of
the
compound of Formula IV will be understood by those skilled in the art. The
compound
capable of ionizing a hydroxyl moiety is preferably a strong base. More
preferably, the
compound capable of ionizing a hydroxyl moiety is selected from the group
consisting of
sodium hydride, potassium hydride, sodium t-butoxide, potassium t-butoxide,
butyl lithium
(BuLi), and lithium disopropylamine. The compound capable of ionizing a
hydroxyl moiety
is more preferably sodium hydride.
The molar ratio of the compound capable of ionizing a hydroxyl moiety on the
PEG
moiety of the compound of Formula IV to the compound of Formula IV is
preferably at least
8
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
about 1:1, and is more preferably at least about 2:1. By providing an excess
of the compound
capable of ionizing the hydroxyl moiety, it is assured that substantially all
of the compounds
of Formula IV are reacted to provide the compounds of Formula I. Thus,
separation
difficulties, which may occur if both compounds of Formula IV and compounds of
Formula I
were present in the reaction product mixture, may be avoided.
Reaction 2 is preferably performed between about 0 C and about 40 C, is more
preferably performed between about 0 C and about 35 C, and is most preferably
performed
between about 0 C and room temperature (approximately 25 C).
Reaction 2 may be performed for various periods of time as will be understood
by
those skilled in the art. Reaction 2 is preferably performed for a period of
time between
about 0.25, 0.5 or 0.75 hours and about 2, 4 or 8 hours.
Reaction 2 is preferably carried out in an aprotic solvent such as, but not
limited to,
N,N-dimethylacetamide (DMA), N,N-dimethylformamide (DMF), dimethyl sulfoxide,
hexamethylphosphoric triamide, tetrahydrofuran (THF), dioxane, diethyl ether,
methyl t-butyl
ether (MTBE), toluene, benzene, hexane, pentane, N-methylpyrollidinone,
dichloromethane,
chloroform, tetrahydronaphthalene, decahydronaphthalene, 1,2-dichlorobenzene,
1,3-
dimethyl-2-imidazolidinone, or a mixture thereof. More preferably, the solvent
is DMF,
dichloromethane or toluene.
Compounds of Formula II are preferably prepared as illustrated in reaction 3:
0
11 30
R2(OC2H4)mOH + CH3SQ R2(OC2H4)mOMs 3
I I
('S') O (II)
R2 and Ms are as described above and the compound of Formula V is present as a
substantially monodispersed mixture of compounds of Formula V; preferably at
least 96, 97,
98 or 99 percent of the compounds in the mixture of compounds of Formula V
have the same
molecular weight; and, more preferably, the mixture of compounds of Formula V
is a
monodispersed mixture.
Q is a halide, preferably chloride or fluoride.
CH3S(02)Q is methanesulfonyl halide. The methanesulfonyl halide is preferably
methanesulfonyl chloride or methanesulfonyl fluoride. More preferably, the
methanesulfonyl
halide is methanesulfonyl chloride.
The molar ratio of the methane sulfonyl halide to the compound of Formula V is
9
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
preferably greater than about 1:1, and is more preferably at least about 2:1.
By providing an
excess of the methane sulfonyl halide, it is assured that substantially all of
the compounds of
Formula V are reacted to provide the compounds of Formula II. Thus, separation
difficulties,
which may occur if both compounds of Formula V and compounds of Formula II
were
present in the reaction product mixture, may be avoided.
Reaction 3 is preferably performed between about -10 C and about 40 C, is more
preferably performed between about 0 C and about 35 C, and is most preferably
performed
between about 0 C and room temperature (approximately 25 C).
Reaction 3 may be performed for various periods of time as will be understood
by
those skilled in the art. Reaction 3 is preferably performed for a period of
time between
about 0.25, 0.5 or 0.75 hours and about 2, 4 or 8 hours.
Reaction 3 is preferably carried out in the presence of an aliphatic amine
including,
but not limited to, monomethylamine, dimethylamine, trimethylamine,
monoethylamine,
diethylamine, triethylamine, monoisopropylamine, diisopropylamine, mono-n-
butylamine, di-
n-butylamine, tri-n-butylamine, monocyclohexylamine, dicyclohexylamine, or
mixtures
thereof. More preferably, the aliphatic amine is a tertiary amine such as
triethylamine.
As will be understood by those skilled in the art, various substantially
monodispersed
mixtures of compounds of Formula V are commercially available. For example,
when R2 is
H or methyl, the compounds of Formula V are PEG or mPEG compounds,
respectively,
which are commercially available from Aldrich of Milwaukee, Wisconsin; Fluka
of
Switzerland, and/or TCl America of Portland, Oregon.
When R2 is a lipophilic moiety such as, for example, higher alkyl, fatty acid,
an ester
of a fatty acid, cholesteryl, or adamantyl, the compounds of Formula V may be
provided by
various methods as will be understood by those skilled in the art. The
compounds of Formula
V are preferably provided as follows:
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
R? OMs + R3(OC2H4)m O-X2+ > R3(OC2H4)m OR2 4
(VI) (VII) (VIII)
R3(OC2H4)m OR2 30 H(OC2H4)m OR2 5
(VIII) (V)
R2 is a lipophilic moiety, preferably higher alkyl, fatty acid ester,
cholesteryl, or
adamantyl, more preferably a lower alkyl ester of a fatty acid, and most
preferably an ethyl
ester of a fatty acid having from 1 to 18 carbon atoms.
R3 is H, benzyl, trityl, tetrahydropyran, or other alcohol protecting groups
as will be
understood by those skilled in the art.
X2+ is a positive ion as described above with respect to X+.
The value of in is as described above.
Regarding reaction 4, a mixture of compounds of Formula VI is reacted with a
mixture of compounds of Formula VII under reaction conditions similar to those
described
above with reference to reaction 1. The mixture of compounds of Formula VI is
a
substantially monodispersed mixture. Preferably, at least 96, 97, 98 or 99
percent of the
compounds in the mixture of compounds of Formula VI have the same molecular
weight.
More preferably, the mixture of compounds of Formula VI is a monodispersed
mixture. The
mixture of compounds of Formula VII is a substantially monodispersed mixture.
Preferably,
at least 96, 97, 98 or 99 percent of the compounds in the mixture of compounds
of Formula
VII have the same molecular weight. More preferably, the mixture of compounds
of Formula
VII is a monodispersed mixture.
Regarding reaction 5, the compound of Formula VIII may be hydrolyzed to
convert
the R3 moiety into an alcohol by various methods as will be understood by
those skilled in the
art. When R3 is benzyl or trityl, the hydrolysis is preferably performed
utilizing H2 in the
presence of a palladium-charcoal catalyst as is known by those skilled in the
art. Of course,
when R3 is H, reaction 5 is unnecessary.
11
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
The compound of Formula VI may be commercially available or be provided as
described above with reference to reaction 3. The compound of Formula VII may
be
provided as described above with reference to reaction 2.
Substantially monodispersed mixtures of polymers comprising PEG moieties and
having the structure of Formula III above can further be reacted with other
substantially
monodispersed polymers comprising PEG moieties in order to extend the PEG
chain. For
example, the following scheme may be employed:
O
I I
R2(OC2H4)m+n-OR' + CH3SQ R2(OC2H4)n,+n-OMs
(III) O (IX)
R2(OC2H4)m+n-OMs + R4(OC2H4)p-O-X2+ R2(OC2H4)m+n+p-OR4
(IX) (X) (XI)
Ms, in and n are as described above with reference to reaction 1; p is similar
to n and
in, and X2+ is similar to X+ as described above with reference to reaction 1.
Q is as described
above with reference to reaction 3. R2 is as described above with reference to
reaction 1 and
is preferably lower alkyl. R' is H. Reaction 6 is preferably performed in a
manner similar to
that described above with reference to reaction 3. Reaction 7 is preferably
performed in a
manner similar to that described above with reference to reaction 1.
Preferably, at least 96,
97, 98 or 99 percent of the compounds in the mixture of compounds of Formula
III have the
same molecular weight, and, more preferably, the mixture of compounds of
Formula III is a
monodispersed mixture. The mixture of compounds of Formula X is a
substantially
monodispersed mixture. Preferably, at least 96, 97, 98 or 99 percent of the
compounds in the
mixture of compounds of Formula X have the same molecular weight, and, more
preferably,
the mixture of compounds of Formula X is a monodispersed mixture.
An embodiment of a method according to the present invention is illustrated by
the
scheme shown in Figure 1, which will now be described. The synthesis of a
substantially
monodispersed mixture of polyethylene glycol-containing oligomers begins by
the
preparation of the monobenzyl ether (XII) of a substantially monodispersed
mixture of
polyethylene glycol. An excess of a commercially available substantially
monodispersed
mixture of polyethylene glycol is reacted with benzyl chloride in the presence
of aqueous
12
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
sodium hydroxide as described by Coudert et al (Synthetic Communications,
16(1): 19-26
(1986)). The sodium salt of XII is then prepared by the addition of NaH, and
this sodium
salt is allowed to react with the mesylate synthesized from the ester of a
hydroxyalkanoic
acid (XIII). The product (XIV) of the displacement of the mesylate is
debenzylated via
catalytic hydrogenation to obtain the alcohol (XV). The mesylate (XVI) of this
alcohol may
be prepared by addition of methanesulfonyl chloride and used as the
electrophile in the
reaction with the sodium salt of the monomethyl ether of a substantially
monodispersed
mixture of a polyethylene glycol derivative, thereby extending the
polyethylene glycol
portion of the oligomer to the desired length, obtaining the elongated ester
(XVII). The ester
may be hydrolyzed to the acid (XVIII) in aqueous base and transformed into the
activated
ester (XIX) by reaction with a carbodiimide and N-hydroxysuccinimide. While
the oligomer
illustrated in Figure 1 is activated using N-hydroxysuccinimide, it is to be
understood that
various other reagents may be used to activate oligomers of the present
invention including,
but not limited to, active phenyl chloroformates such as para-nitrophenyl
chloroformate,
phenyl chloroformate, 3,4-phenyldichloroformate, and 3,4-
phenyldichloroformate;
tresylation; and acetal formation.
Still referring to Figure 1, q is from 1 to 24. Preferably, q is from 1 to 18,
and q is
more preferably from 4 to 16. R4 is a moiety capable of undergoing hydrolysis
to provide the
carboxylic acid. R4 is preferably lower alkyl and is more preferably ethyl.
The variables n
and in are as described above with reference to reaction 1.
All starting materials used in the procedures described herein are either
commercially
available or can be prepared by methods known in the art using commercially
available
starting materials.
The present invention will now be described with reference to the following
examples. It should be appreciated that these examples are for the purposes of
illustrating
aspects of the present invention, and do not limit the scope of the invention
as defined by the
claims.
EXAMPLES
Examples I through 6 refer to the scheme illustrated in Figure 2.
13
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
Example 1
Hexaethylene glycol monobenzyl ether (XX)
An aqueous sodium hydroxide solution prepared by dissolving 3.99 g (100 mmol)
NaOH in 4 ml water was added slowly to non-polydispersed hexaethylene glycol
(28.175 g,
25 ml, 100 mmol). Benzyl chloride (3.9 g, 30.8 mmol, 3.54 ml) was added and
the reaction
mixture was heated with stirring to 100 C for 18 hours. The reaction mixture
was then
cooled, diluted with brine (250 ml) and extracted with methylene chloride (200
ml x 2). The
combined organic layers were washed with brine once, dried over Na2SO4,
filtered and
concentrated in vacuo to a dark brown oil. The crude product mixture was
purified via flash
chromatography (silica gel, gradient elution: ethyl acetate to 9/1 ethyl
acetate/methanol) to
yield 8.099 g (70 %) of non-polydispersed XX as a yellow oil.
Example 2
Ethyl 6-methylsulfonyloxyhexanoate (XXI)
A solution of non-polydispersed ethyl 6-hydroxyhexanoate (50.76 ml, 50.41 g,
227
mmol) in dry dichloromethane (75 ml) was chilled in a ice bath and placed
under a nitrogen
atmosphere. Triethylamine (34.43 ml, 24.99 g, 247 mmol) was added. A solution
of
methanesulfonyl chloride (19.15 ml, 28.3 g, 247 mmol) in dry dichloromethane
(75 ml) was
added dropwise from an addition funnel. The mixture was stirred for three and
one half
hours, slowly being allowed to come to room temperature as the ice bath
melted. The
mixture was filtered through silica gel, and the filtrate was washed
successively with water,
saturated NaHCO3, water and brine. The organics were dried over Na2SO4,
filtered and
concentrated in vacuo to a pale yellow oil. Final purification of the crude
product was
achieved by flash chromatography (silica gel, 1/1 hexanes/ethyl acetate) to
give the non-
polydispersed product (46.13 g, 85 %) as a clear, colorless oil. FAB MS: m/e
239 (M+H),
193 (M-CZH50).
Example 3
6-{2-[2-(2-{2-[2-(2-Benzyloxyethoxy)ethoxy] ethoxy}-ethoxy)-
ethoxy]-ethoxy}-hexanoic acid ethyl ester (XXII)
Sodium hydride (3.225 g or a 60 % oil dispersion, 80.6 mmol) was suspended in
80
ml of anhydrous toluene, placed under a nitrogen atmosphere and cooled in an
ice bath. A
14
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
solution of the non-polydispersed alcohol XX (27.3 g, 73.3 mmol) in 80 ml dry
toluene was
added to the NaH suspension. The mixture was stirred at 0 C for thirty
minutes, allowed to
come to room temperature and stirred for another five hours, during which time
the mixture
became a clear brown solution. The non-polydispersed mesylate XXI (19.21 g,
80.6 mmol)
in 80 ml dry toluene was added to the NaH/alcohol mixture, and the combined
solutions were
stirred at room temperature for three days. The reaction mixture was quenched
with 50 ml
methanol and filtered through basic alumina. The filtrate was concentrated in
vacuo and
purified by flash chromatography (silica gel, gradient elution: 3/1 ethyl
acetate/hexanes to
ethyl acetate) to yield the non-polydispersed product as a pale yellow oil
(16.52 g, 44 %).
FAB MS: m/e 515 (M+H).
Example 3
6-{2-[2-(2-{2-[2-(2-hydroxyethoxy)ethoxy] ethoxy}-ethoxy)-
ethoxy]-ethoxy}-hexanoic acid ethyl ester (XXIII)
Non-polydispersed benzyl ether XI (1.03 g, 2.0 mmol) was dissolved in 25 ml
ethanol. To this solution was added 270 mg 10 % Pd/C, and the mixture was
placed under a
hydrogen atmosphere and stirred for four hours, at which time TLC showed the
complete
disappearance of the starting material. The reaction mixture was filtered
through Celite 545
to remove the catalyst, and the filtrate was concentrated in vacuo to yield
the non-
polydispersed title compound as a clear oil (0.67 g, 79 %). FAB MS: m/e 425
(M+H), 447
(M+Na).
Example 4
6-{2-[2-(2-{2-[2-(2-methylsulfonylethoxy)ethoxy] ethoxy}-
ethoxy)-ethoxy]-ethoxy}-hexanoic acid ethyl ester (XXIV)
The non-polydispersed alcohol XXIII (0.835 g, 1.97 mmol) was dissolved in 3.5
ml
dry dichloromethane and placed under a nitrogen atmosphere. Triethylamine
(0.301 ml,
0.219 g, 2.16 mmol) was added and the mixture was chilled in an ice bath.
After two
minutes, the methanesulfonyl chloride (0.16 ml, 0.248 g, 2.16 mmol) was added.
The
mixture was stirred for 15 minutes at 0 C, then at room temperature for two
hours. The
reaction mixture was filtered through silica gel to remove the
triethylammonium chloride,
and the filtrate was washed successively with water, saturated NaHCO3, water
and brine.
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
The organics were dried over Na2SO4, filtered and concentrated in vacuo. The
residue was
purified by column chromatography (silica gel, 9/1 ethyl acetate/methanol) to
give non-
polydispersed XXIV as a clear oil (0.819 g, 83 %). FAB MS: m/e 503 (M+H).
Example 5
8-[2-(2-{2-[2-(2-{2-[2-(2-methoxyethoxy)ethoxy]-ethoxy}-ethoxy)-
ethoxy]-ethoxy}-ethoxy)-ethoxy]-hexanoic acid ethyl ester (XXV)
NaH (88 mg of a 60 % dispersion in oil, 2.2 mmol) was suspended in anhydrous
toluene (3 ml) under N2 and chilled to 0 OC. Non-polydispersed diethylene
glycol
monomethyl ether (0.26 ml, 0.26 g, 2.2 mmol) that had been dried via
azeotropic distillation
with toluene was added. The reaction mixture was allowed to warm to room
temperature and
stirred for four hours, during which time the cloudy grey suspension became
clear and yellow
and then turned brown. Non-polydispersed mesylate XXIV (0.50 g, 1.0 mmol) in
2.5 ml dry
toluene was added. After stirring at room temperature over night, the reaction
was quenched
by the addition of 2 m] of methanol and the resultant solution was filtered
through silica gel.
The filtrate was concentrated in vacuo and the FAB MS: m/e 499 (M+H), 521
(M+Na).
Additional purification by preparatory chromatography (silica gel, 19/3
chloroform/methanol) provided the non-polydispersed product as a clear yellow
oil (0.302 g
57 %). FAB MS: m/e 527 (M+H), 549 (M+Na).
Example 6
8-[2-(2-{2-[2-(2-{2-[2-(2-methoxyethoxy)ethoxy]-ethoxy}-
ethoxy)-ethoxy]-ethoxy}-ethoxy)-ethoxy]-hexanoic acid (XXVI)
Non-polydispersed ester XXV (0.25 g, 0.46 mmol) was stirred for 18 hours in
0.71 ml
of 1 N NaOH. After 18 hours, the mixture was concentrated in vacuo to remove
the alcohol
and the residue dissolved in a further 10 ml of water. The aqueous solution
was acidified to
pH 2 with 2 N HCI and the product was extracted into dichloromethane (30 ml x
2). The
combined organics were then washed with brine (25 ml x 2), dried over Na2SO4,
filtered and
concentrated in vacuo to yield the non-polydispersed title compound as a
yellow oil (0.147 g,
62 %). FAB MS: m/e 499 (M+H), 521 (M+Na).
16
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
Example 7
8-[2-(2-{2-[2-(2-{2-[2-(2-methoxyethoxy)ethoxy]-ethoxy}-ethoxy)-ethoxy]-
ethoxy}-
ethoxy)-ethoxy]-hexanoic acid 2,5-dioxo-pyrrolidin-l-yl ester (XXVII)
Non-polydispersed acid XXVI (0.209 g, 0.42 mmol) were dissolved in 4 ml of dry
dichloromethane and added to a dry flask already containing NHS (N-
hydroxysuccinimide)
(57.8 mg, 0.502 mmol) and EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride) (98.0 mg, 0.502 mmol) under a N2 atmosphere. The solution was
stirred at
room temperature overnight and filtered through silica gel to remove excess
reagents and the
urea formed from the EDC. The filtrate was concentrated in vacuo to provide
the non-
polydispersed product as a dark yellow oil (0.235 g, 94 %). FAB MS: m/e 596
(M+H), 618
(M+Na).
Examples 8 through 17
Reactions in Examples 8 through 17 were carried out under nitrogen with
magnetic
stirring, unless otherwise specified. "Work-up" denotes extraction with an
organic solvent,
washing of the organic phase with saturated NaCl solution, drying (MgSO4), and
evaporation
(rotary evaporator). Thin layer chromatography was conducted with Merck glass
plates
precoated with silica gel 60 F - 254 and spots were visualized by iodine
vapor. All mass
spectra were determined by Macromolecular Resources Colorado State University,
CO and
are reported in the order m/z, (relative intensity). Elemental analyses and
melting points were
performed by Galbraith Laboratories, Inc., Knoxville, TN. Examples 8-17 refer
to the
scheme illustrated in Figure 3.
Example 8
8-Methoxy-l-(methylsulfonyl)oxy-3,6-dioxaoctane (XXXI)
A solution of non-polydispersed triethylene glycol monomethyl ether molecules
(4.00
mL, 4.19 g, 25.5 mmol) and triethylamine (4.26 mL, 3.09 g, 30.6 mmol) in dry
dichloromethane (50 mL) was chilled in an ice bath and place under a nitrogen
atmosphere.
A solution of methanesulfonyl chloride (2.37 mL, 3.51 g, 30.6 mmol) in dry
dichloromethane
(20 mL) was added dropwise from an addition funnel. Ten minutes after the
completion of
the chloride addition, the reaction mixture was removed from the ice bath and
allowed to
come to room temperature. The mixture was stirred for an additional hour, at
which time
17
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
TLC (CHC13 with 15% MeOH as the elutant) showed no remaining triethylene
glycol
monomethyl ether.
The reaction mixture was diluted with another 75 mL of dichloromethane and
washed
successively with saturated NaHCO3i water and brine. The organics were dried
over Na2SO4,
filtered and concentrated in vacuo to give non-polydispersed compound XXXI as
a clear oil
(5.31 g, 86%).
Example 9
Ethylene glycol mono methyl ether (XXXII) (m=4,5,6)
To a stirred solution of non-polydispersed compound XXVIII (35.7 mmol) in dry
DMF (25.7 mL), under N2 was added in portion a 60% dispersion of NaH in
mineral oil, and
the mixture was stirred at room temperature for 1 hour. To this salt XXIX was
added a
solution of non-polydispersed mesylate XXXI (23.36) in dry DMF (4 ml) in a
single portion,
and the mixture was stirred at room temperature for 3.5 hours. Progress of the
reaction was
monitored by TLC (12% CH3OH-CHC13). The reaction mixture was diluted with an
equal
amount of IN HCI, and extracted with ethyl acetate (2 x 20 ml) and discarded.
Extraction of
aqueous solution and work-up gave non-polydispersed polymer XXXII (82 -84%
yield).
Example 10
3,6,9,12,15,18,21-Heptaoxadocosanol (XXXII) (m=4)
Oil; Rf 0.46 (methanol : chloroform = 3:22); MS m/z calc'd for C15H3208 340.21
(M++l), found 341.2.
Example 11
3,6,9,12,15,18,21,24-Octaoxapentacosanol (XXXII) (m=5)
Oil; Rf 0.43 (methanol : chloroform = 6:10); MS m/z calc'd for C17H3609 384.24
(M++1), found 385.3.
Example 12
3,6,9,12,15,18,21,24,27-Nonaoxaoctacosanol (XXXII) (m=5)
Oil; Rf 0.42 (methanol : chloroform = 6:10); MS m/z calc'd for C19H40O10
428.26
(M++1), found 429.3.
18
CA 02449698 2003-12-04
WO 02/098949 PCT/US02/17619
Example 13
20-methoxy-l-(methylsulfonyl)oxy-3,6,9,12,15,18-hexaoxaeicosane (XXXIII)
Non-polydispersed compound XXXIII was obtained in quantitative yield from the
alcohol XXXII (m=4) and methanesulfonyl chloride as described for XXXI, as an
oil; Rf 0.4
(ethyl acetate : acetonitrile = 1:5); MS m/z calc'd for C17H37O10 433.21
(M++1), found
433.469.
Example 14
Ethylene glycol mono methyl ether (XXXIV) (m=3,4,5)
The non-polydispersed compounds XXXIV were prepared from a diol by using the
procedure described above for compound XXXII.
Example 15
3,6,9,12,15,18,21,24,27,30-Decaoxaheneicosanol (XXXIV) (m=3)
Oil; Rf 0.41 (methanol : chloroform = 6:10); MS m/z calc'd for C21H44011
472.29
(M++1), found 472.29.
Example 16
3,6,9,12,15,18,21,24,27,30,33-Unecaoxatetratricosanol (XXXIV) (m=4)
Oil; Rf 0.41 (methanol : chloroform = 6:10); MS m/z calc'd for C23H48012
516.31
(M++1), found 516.31.
Example 17
3,6,9,12,15,18,21,24,27,30,33,36-Dodecaoxaheptatricosanol (XXXIV) (m=5)
Oil; Rf 0.41 (methanol : chloroform = 6:10); MS m/z calc'd for C25H52013
560.67
(M++1), found 560.67.
In the specification, there has been disclosed typical preferred embodiments
of the
invention and, although specific terms are employed, they are used in a
generic and
descriptive sense only and not for purposes of limitation, the scope of the
invention being set
forth in the following claims.
19
